cgb2 And cgb1 Genes; Diagnosis, Monitoring And Treatment Of Cancer

Abstract

The present invention relates to methods for screening human tissue or fluid samples for changes in the expression of genes, in particular CGB2 and CBG1, characteristic of poor prognosis in cancer such as bladder cancer. The methods have application for the screening, diagnosis or monitoring of other common epithelial cancers including those of the bladder, breast, cervix, colon, endometrium, kidney, lung (including small cell lung carcinoma—SCLC), nasal/pharynx, oro/facial, ovary, prostate, pancreas, vagina and vulva. Methods of treating cancers by reducing the level of expression of CGB2 and CGB1 or products thereof are also disclosed.

1 . A method for investigating an epithelial cancer, the method comprising detecting and/or measuring the expression product of the CGB2 and/or CGB1 gene(s). 
     
     
         2 . A method according to  claim 1  wherein the expression product of the CGB2 and/or CGB1 gene is the 230 bp splice variant of the CGB2 and/or CGB1 gene(s). 
     
     
         3 . A method according to  claim 1 , further comprising detecting and/or measuring the expression of any one or more of the genes CGB3, CGB5, CGB7, and/or CGB8. 
     
     
         4 . A method according to  claim 1 , wherein detecting and/or measuring the expression product of the CGB2 and/or CGB1 gene(s) comprises detecting the levels of mRNA, cDNA, peptide or protein. 
     
     
         5 . A method according to  claim 1 , wherein detecting and/or measuring the expression product of the CGB2 and/or CGB1 gene(s) comprises detecting the levels of mRNA or cDNA. 
     
     
         6 . A method according to  claim 5  wherein the method utilises a primer which hybridises to a sequence within Exon 1 of the CGB2 and/or CGB1 gene and/or a reverse primer which hybridises to a sequence after the initiation site in Exon 2 of the CGB2 and/or CGB1 gene. 
     
     
         7 . A method according to  claim 6  wherein the forward primer hybridises to a sequence at the Exon1/Exon2 boundary the CGB2 and/or CGB1 gene. 
     
     
         8 . A method according to  claim 6  wherein the forward primer is 5′-CGTCCAACACCCCTCACTCC-3′ (SEQ ID NO:4) and/or the reverse primer is 5′-5 GGCAGCCCTCCTTCTCCAC-3′ (SEQ ID. NO:5 SEQ ID NO:5). 
     
     
         9 . A method according to  claim 5  further comprising utilising the forward primer 5′-CAGCACCTTTCTCGGGTCAC-3′ (SEQ ID NO:6) and/or the reverse primer 5′-CAGGGAGTAGGGTGTAGGAAGG-3′ (SEQ ID NO:7). 
     
     
         10 . A method according to  claim 5 , wherein the method employs nucleic acid hybridisation for the amplification of CGB genes. 
     
     
         11 . A method according to  claim 5 , utilising a nucleic acid probe capable of hybridising to the nucleic acid expression product of the CGB2 and/or CGB1 gene(s). 
     
     
         12 . A method according to  claim 11 , wherein the nucleic acid probe forms part of an array. 
     
     
         13 . A method according to  claim 1 , wherein detecting and/or measuring the expression product of the CGB2 and/or CGB1 gene(s) comprises detecting the levels of the peptide, protein or pre-protein. 
     
     
         14 . A method according to  claim 13  wherein the peptide, protein or pre-protein comprises SEQ ID NO:2. 
     
     
         15 . A method of  claim 1 , wherein said method is carried out on a sample obtained from a subject. 
     
     
         16 . A method of  claim 15 , wherein said sample is selected from a blood, urine, cerebrospinal fluid, sputum, bronchial lavage, saliva or tissue sample. 
     
     
         17 . A method of  claim 15 , wherein said sample comprises circulating tumour cells and/or stem cells isolated from the blood of a subject. 
     
     
         18 . A kit of parts for detecting and/or measuring the expression of the CGB2 and/or CGB1 gene(s) comprising an agent for detecting and/or measuring the expression product of the CGB2 and/or CGB1 gene(s). 
     
     
         19 . The kit of  claim 18 , wherein the agent for detecting and/or measuring the expression of the CGB2 and/or CGB1 gene(s) is selected from any one or more of any one or more of
 (a) a set of primers capable of hybridising to the 230 bp splice variant transcript,   (b) a probe capable of hybridizing to the 230 bp splice variant transcript, and   (c) an antibody capable of specifically binding to the pre-protein or protein translated from the 230 bp splice variant transcript.   
     
     
         20 . The kit of  claim 19  further comprising primers or hybridisation probes capable of hybridising with a housekeeping gene, used as a control. 
     
     
         21 . A method of treating an hCGβ secreting cancerous condition comprising administering to patient in need thereof, a pharmaceutically acceptable amount of an agent capable of reducing the level of secreted hCGβ. 
     
     
         22 . The method of  claim 21 , wherein the agent capable of reducing the level of secreted hCGβ is an agent that reduces the expression of one or more CGB genes. 
     
     
         23 . The method of  claim 21  wherein the agent is siRNA molecules which specifically bind to sequence of the CGB genes. 
     
     
         24 - 29 . (canceled) 
     
     
         30 . An expression product of CGB1 or CGB2. 
     
     
         31 . The expression product of  claim 30  wherein the expression product is the 230 bp splice variant transcript. 
     
     
         32 . The expression product of  claim 30  wherein the expression product comprises a nucleic acid selected from
 (a) A nucleic acid of SEQ. ID NO:1; 
 (b) A nucleic acid sequence encoding the polypeptide sequence of SEQ ID NO:2 or SEQ ID NO:15; 
 (c) A sequence substantially identical to the sequence of (a) or (b); or a homologue or variant thereof; and 
 (d) A sequence complementary or capable of hybridising to a sequence of any one of (a) to (c). 
 
     
     
         33 . The expression product of  claim 30  wherein the expression product is an amino acid sequence. 
     
     
         34 . The expression product of  claim 30  wherein the expression product is a polypeptide of SEQ ID NO:2 or SEQ ID NO:15 or a substantially identical polypeptide thereof, or a homologue or derivative thereof.